tbuckley/cabot-search huggingface dataset:
Abstract 3937: An intracellular variant of zona pellucida glycoprotein 3 is expressed in cancer
Authors: Iman J. Schultz, Yvette Zimmerman, Cathy B. Moelans, Marcin Chrusciel, Jan Krijgh, Paul J. van Diest, Ilpo T. Huhtaniemi, Herjan J. Coelingh Bennink
Journal: Cancer Research (2023)

This study describes the expression of an intracellular variant of the Zona Pellucida glycoprotein 3 (ZP3) that is predominantly expressed in cancer. ZP3 is an important constituent of the Zona Pellucida (ZP), the extracellular layer surrounding oocytes. It is highly expressed in maturing oocytes, but not in other adult human tissues. As ZP3 protein has also been detected in cancer, it may serve as an interesting diagnostic and therapeutic target. Here, we extend the cancer-related ZP3 expression data using immunohistochemistry (IHC) of tumor biopsies, interrogating publicly available RNA-sequencing (RNA-seq) data of cancer cell lines (CCLs) and tumor and normal tissues, as well as computational analysis and real-time quantitative PCR (qPCR) of CCLs. IHC data for several cancer types shows abundant ZP3 protein expression, which is confined to the cytoplasm, contradicting the extracellular ZP3 localization in oocytes. An alternative ZP3 mRNA variant, which we term 'ZP3-Cancer', is annotated in the NCBI and Ensembl databases, and lacks the genetic information encoding the N-terminal signal peptide that governs extracellular secretion. Analysis of publicly available RNA-seq data of 1339 CCLs indicates that ZP3-Cancer is the dominant variant as compared to ZP3-Oocyte, which was validated by independent computational analysis. Expression of ZP3-Cancer in CCLs was confirmed by qPCR. Publicly available RNAseq data of tumor and normal tissues confirms strongly enhanced expression of ZP3-Cancer in cancer cells. In addition, ZP3-Cancer expression is upregulated in advanced stages and shows a relationship with patient survival in a number of cancer types. The folliculogenesis specific transcription factor FIGLA, which activates transcription of ZP3-Oocyte in maturing oocytes, appears absent in cancer as inferred from publicly available transcriptome data, indicating alternative ZP3-Cancer transcriptional activation mechanisms in this disease. The cancer restricted expression of this ZP3 transcript variant renders it an attractive tumor specific antigen for the development of a therapeutic cancer vaccine, particularly using mRNA technology.Citation Format: Iman J. Schultz, Yvette Zimmerman, Cathy B. Moelans, Marcin Chrusciel, Jan Krijgh, Paul J. van Diest, Ilpo T. Huhtaniemi, Herjan J. Coelingh Bennink. An intracellular variant of zona pellucida glycoprotein 3 is expressed in cancer. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3937.
Abstract 2872: Site specific tumor microenvironments modulate the immune checkpoint inhibitors response in models of CT26 colon carcinoma
Authors: Fei Zhao, Caitlin Thompson, Nicole Barnes, José Roques, Terrence Coleman, Tyler Rowe, Joseph Kolb, Chassidy Hall, Patrick Fadden
Journal: Cancer Research (2023)

Abstract Syngeneic models provide a robust platform to test new and cutting edge immune-oncology therapies. CT26 is a popular syngeneic colon carcinoma model that is known to respond to immune checkpoint inhibitors when engrafted into Balb/c mice as a subcutaneous flank tumor. The subcutaneous flank implant location provides a robust model, that is easy to set up, and allows for continuous measurement of the tumor growth by calipering. However, the location of tumor implant has been shown to impact tumor microenvironment and immune response for a number of cancer cell lines (Zhao 2017, Brand 2021, Oliver 2018). In addition to subcutaneous flank tumors, CT26 can be implanted to generate a lung metastasis or peritoneal metastasis model. We were interested to see if the lung metastasis or peritoneal metastasis CT26 model responded differently to immune checkpoint inhibition compared with the subcutaneous model and whether the different implant locations resulted in changes to the tumor infiltrating lymphocytes. CT26 cells were either injected intravenously to induce a lung metastasis model, intraperitoneally to produce a peritoneal metastasis model, or subcutaneously to generate a solid flank tumor model. Immune checkpoint inhibitor (ICI) therapy began on Day 5 following implant in each model, and tumor progression or survival provided the endpoint for analysis. In addition, tumor infiltrating leukocytes were assessed by flow cytometry to determine differences in the overall tumor microenvironment for each model. The response to both anti-PD-1 and anti-CTLA-4 monotherapies were strongest in the subcutaneous flank model, followed by moderate responses in the lung metastasis model, and no significant response to either ICI in the peritoneal metastasis model. This provides researchers with an opportunity to assess the ability of novel immune-oncology therapies to provoke an immunogenic response in models with identical antigenicity, but different tumor microenvironments based on the tumor location. Citation Format: Fei Zhao, Caitlin Thompson, Nicole Barnes, José Roques, Terrence Coleman, Tyler Rowe, Joseph Kolb, Chassidy Hall, Patrick Fadden. Site specific tumor microenvironments modulate the immune checkpoint inhibitors response in models of CT26 colon carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2872.
Abstract 1018: Multiplex immunofluorescence and image analysis to investigate the role of the immune contexture and fibroblast activation for tumor cell budding in colorectal cancer
Authors: Dirk Zielinski, Rebekka Vogtmann, Anne Siemon, Christina Koppel, Christina Schipper, Anastasiia Tereshchenko, Suso Platero, Hans-Ulrich Schildhaus
Journal: Cancer Research (2023)

Abstract The advent of immune oncology had a significant impact on the stratification of cancer patients in the past few years. Furthermore, immune cell phenotyping of the tumor microenvironment is becoming a tool not only for the identification of novel predictive biomarkers for cancer immunotherapy, but also for prognostic markers that may help to understand several mechanisms like invasion and epithelial-mesenchymal transition (EMT). We demonstrate how multiplexed immunofluorescence (mIF) assays and digital image analysis helped to investigate tumor budding in colorectal cancer by evaluating the tumor microenvironment and activated fibroblasts. Akoya Bioscience’s Opal detection system and fluorophores were optimized on LEICA Bond RX for use with human normal tonsil control tissues and FFPE colorectal cancer specimens. The 6-plex mIF panel included CD4 (clone SP35), CD8 (clone C8/144B), CD68 (clone PG-M1), FoxP3 (clone SP97), PD-L1 (clone SP263) and pan-Cytokeratin (clones AE1/AE3). In addition, FAP-alpha single-plex IHC was used to investigate reactivity of the stroma surrounding tumor cell buds. Each single marker of the mIF panel was independently validated for specificity regarding stripping efficacy and epitope stability, and for accuracy compared to single plex bright field immunohistochemistry. Automated image analysis and cellular phenotyping followed a workflow of custom Visiopharm apps. The tissue was segmented into regions of interest, such as areas with and without tumor cell budding, and bud microenvironment. Additional parameters like MSI (Microsatellite Instability) status and TNM classification were also taken into consideration for data analysis. The applied mIF immune cell panel and automated image analysis workflow identified a broad number of different immune cell phenotypes, including rare double- or triple-positive cell subtypes, as well as their spatial relation to tumor cell buds within the activated stroma region. This revealed new insights into the complexity of the tumor microenvironment in colorectal cancer around tumor cell buds, suggesting a potential effect of both the immune system and the stromal cells on tumor cell invasion into the surrounding tissue. Citation Format: Dirk Zielinski, Rebekka Vogtmann, Anne Siemon, Christina Koppel, Christina Schipper, Anastasiia Tereshchenko, Suso Platero, Hans-Ulrich Schildhaus. Multiplex immunofluorescence and image analysis to investigate the role of the immune contexture and fibroblast activation for tumor cell budding in colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1018.
Abstract 5015: Wee1 inhibition and its synthetic lethal combination with Chk1 inhibition in mouse model of neuroendocrine prostate cancer
Authors: Yapeng Chao, Yuzhou Chen, Yu Liu, Jaclyn A. Connelly, Hong Wang, Qiming Jane Wang
Journal: Cancer Research (2023)

Abstract The mitotic regulator Wee1 is a critical regulator of G2/M cell cycle checkpoint and DNA damage response pathways. Wee1-targeted agents are under active clinical investigation in multiple cancer types. However, Wee1 and its inhibitors have not been well studied in prostate cancer, particularly neuroendocrine prostate cancer (NEPC). In this study, we investigated the therapeutic potential of Wee1 inhibitor AZD1775 and its synthetic lethal combination with Chk1 inhibitor in the context of NEPC. Our data demonstrated increased Wee1 expression in NEPC cells and tumors. Treatment of NEPC cells with AZD1775 inhibited NEPC tumor cell survival with greater potency as compared to other prostate cancer cells, demonstrating increased vulnerability of NEPC cells to Wee1 inhibition. We further identified synthetic lethality between the inhibition of Wee1 and several protein kinases, particularly Chk1, another key G2/M checkpoint kinase. Using an autochthonous adenocarcinoma mouse prostate (TRAMP) model that gives rise to NEPC tumors, we assessed the efficacy of AZD1775 and a highly selective Chk1 inhibitor SRA737 as single agents and in combination on the progression, survival, and metastatic spread of NEPC tumors in vivo. AZD1775 and SRA737 when used as single agents significantly suppressed tumor growth, improved overall survival, and incidence of distant metastases of mice with NEPC tumors. The combination of AZD1775 and SRA737 on a sequential 2-wk treatment schedule was more efficacious in suppressing tumor growth and metastasis than each drug alone in castrated TRAMP NEPC tumors. Mechanistically, the combination of Chk1 and Wee1 inhibitors synergized to reduce the inhibitory phosphorylation of CDK1 at Y15 and Cdc25c at S216, resulting in enhanced activation of CDK1/cyclin B complex, abrogation of G2/M checkpoint, and consequent premature mitotic entry, which ultimately led to mitotic catastrophe and apoptosis. Our findings provide promising preclinical evidence supporting the therapeutic values of Wee1 and Chk1 inhibitors as single agents and in combination for the treatment of NEPC. Citation Format: Yapeng Chao, Yuzhou Chen, Yu Liu, Jaclyn A. Connelly, Hong Wang, Qiming Jane Wang. Wee1 inhibition and its synthetic lethal combination with Chk1 inhibition in mouse model of neuroendocrine prostate cancer. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5015.
Abstract 3994: Differential regulation of hormone receptors and Ki67 in luminal breast carcinomas and tumor-adjacent mammary glands by mifepristone treatment
Authors: Leo Saldain, Silvia Vanzulli, Paula Martinez Vazquez, Javier Burruchaga, Eunice Spengler, Claudia Lanari, Paola Rojas
Journal: Cancer Research (2023)

Abstract Since antiprogestins inhibit the growth of preclinical mammary tumor models expressing higher levels of progesterone receptor (PR) isoform A (PRA) than isoform B (PRB), we designed a window-of-opportunity trial (MIPRA; NCT02651844) to study the benefits of mifepristone (MFP) in luminal breast carcinomas from postmenopausal patients selected by their PR+ expression (&amp;gt;50%) and their PR isoform ratio (PRA/PRB&amp;gt;1.5; PRA-H). A decrease in the cell proliferation marker Ki67 was registered after treatment in 14 out of the 20 tumors evaluated. This inhibition was associated with a decrease in PR, estrogen receptor (ER) and pSer118ER evaluated by immunohistochemistry while no changes in pSer167ER expression were observed (PMID: 36269797). In selected tissues from these samples we observed that the staining intensity in trapped glands within the MFP-treated tumors did not follow a similar trend as that of tumor cells. Thus, the aim of this study was to evaluate the expression of hormone receptors and Ki67 in non-neoplastic human mammary glands (nnMG) adjacent to the PRA-H tumor tissue of postmenopausal patients, treated (n=8) or not (n=9) with MFP. No differences in PR, ER and pSer118ER expression were observed between MFP-treated nnMG and those from untreated patients. Notably, contrarily to what occurred in tumors, there was a significant decrease of pSer167ER expression (p=0.008) in the MFP-treated nnMG compared to the untreated glands, suggesting a selective modulation in the AKT-mediated activation of ER. Regarding the proliferative state of the nnMGs, there was a slight but significant increase (p=0.02) in Ki67 expression in MFP-treated vs. non-treated nnMG. Data regarding the effect of antiprogestins in normal mammary glands is limited and points, in premenopausal women, to a decrease in the Ki67 index after treatment. The basal quiescent status of nnMG in postmenopausal women may explain this slight stimulatory effect. In conclusion, our results show that MFP exerts a specific regulatory effect in PRA-H tumors that is not observed in the nnMG, probably due to the fact that nnMG have presumably equimolar levels of PRA and PRB. In addition, to counteract the possible stimulatory effect that MFP may induce on nnMG, the combination of MFP and tamoxifen treatment is suggested. Citation Format: Leo Saldain, Silvia Vanzulli, Paula Martinez Vazquez, Javier Burruchaga, Eunice Spengler, Claudia Lanari, Paola Rojas. Differential regulation of hormone receptors and Ki67 in luminal breast carcinomas and tumor-adjacent mammary glands by mifepristone treatment. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3994.
Abstract 2446: An investigation of HOX-regulatory mechanisms and their role in the SC origin of human CRC
Authors: Brian Osmond, Caroline O. Facey, Lynn M. Opdenaker, Chi Zhang, Bruce M. Boman
Journal: Cancer Research (2023)

Abstract HOX genes are a highly conserved subset of genes that encode for transcription factors critical for stem cell (SC) function and embryonic development. Several HOX genes have also been implicated in a myriad of cancers, including colorectal cancer (CRC). However, the mechanisms by which HOX genes contribute to cancer development are still not fully understood. Specifically, the mechanisms by which HOX genes regulate SC populations and how SC dysregulation leads to tumorigenesis needs to be clarified. Current research indicates that HOX genes are regulated by WNT and retinoic acid (RA) signaling. Goal: To ascertain how HOX-regulatory mechanisms play a role in the SC origin of human CRC. Hypotheses: (1) HOX genes regulated by RA and WNT signaling control SC differentiation along various lineages in normal human colonic epithelium. (2) Aberrant expression of HOX genes leads to decreased differentiation and CSC overpopulation in CRC. Preliminary data shows that RA signaling occurs in ALDH+ SCs and HOXA4, HOXA9 &amp; HOXD10 are selectively expressed in ALDH+ SCs. Moreover, treatment of CRC cells with all-trans retinoic acid (ATRA) modulates expression of HOXA4, HOXA9, HOXC8, HOXC9, &amp; HOXD10 while also reducing cell count. Nanostring profiling also shows that expression of select HOX genes (HOXA3, HOXA4, HOXA5, HOXA10, HOXB6, HOXB8, HOXB9, &amp; HOXD9) significantly changes in response to ATRA. Bioinformatics analysis has identified RA response elements (RAREs) within the promoters of all mentioned HOX genes. Understanding how HOX genes control normal colonic SCs, and, how they become dysregulated in CSCs will identify mechanisms that help explain how HOX genes contribute to SC overpopulation that drives colorectal cancer (CRC) growth. Citation Format: Brian Osmond, Caroline O. Facey, Lynn M. Opdenaker, Chi Zhang, Bruce M. Boman. An investigation of HOX-regulatory mechanisms and their role in the SC origin of human CRC [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2446.
Abstract 678: Investigating the mechanisms involved in HMGB1-dependent DNA uptake and STING activation in dendritic cells
Authors: Daiana P. Celias, Kay Hänggi, Brian Ruffell
Journal: Cancer Research (2023)

Abstract Background: The cGAS-STING pathway is critical for the development of anti-tumor immunity. Activation of host STING can occur by sensing of extracellular DNA by intratumoral dendritic cells (DCs). After chemotherapy and under conditions of chromosomal instability, tumor cells release double-stranded DNA. We have previously shown DCs are able to internalize DNA in a dynamin-dependent manner, suggesting that a membrane receptor might mediate DNA endocytosis. DNA uptake also requires the presence of the high mobility group box protein 1 (HMGB1). HMGB1 has multiple known receptors such as TLR2, TLR4, RAGE and CD24. However, whether these receptors are involved in DNA engulfment by DCs remains unknown. Here, we sought to elucidate the mechanisms facilitating DNA uptake and subsequent STING activation in DCs. Methods: Bone marrow derived (BM)DCs obtained from wild type (WT), RAGE, TLR2 or TLR4-deficient mice, as well as and WT or CD24-deficient MutuDC1940 cells, were cultured in the presence or absence of Cy5-labeled plasmid DNA, a recombinant Flag-tagged HMGB1 protein (rHMGB1) and/or DMXAA. Cy5-DNA uptake, Flag-HMGB1 binding and IRF3 phosphorylation was analyzed by flow cytometry or immunofluorescence microscopy. MutuDC1940 cells were also treated with plasmid DNA, rHMGB1, tumor debris (HS) and/or L-Leucyl-L-Leucine methyl ester (LLOMe) and galectin-3 clustering was quantified by immunofluorescence. Statistically significant differences were determined by ANOVA test. Results: DCs take-up Cy5-DNA when rHMGB1 was added, and it was HMGB1 dose-dependent, supporting HMGB1 as a key factor for DNA uptake. We found that TLR2, TLR4, RAGE or CD24 deficient DCs were capable of internalizing Cy5-DNA in the presence of rHMGB1 to the same extent as WT DCs, indicating that none of these receptors are necessary for HMGB1-mediated DNA uptake. DNA-HMGB1 stimulation triggered IRF3 phosphorylation in DCs, suggesting extracellular DNA enters the cytosol and is sensed by cGAS, leading to STING activation. In support of this, DNA-HMGB1 treatment induced an increase in lysosomal membrane permeabilization, as measured by the extent of galectin-3 clusters within cells. Conclusion: HMGB1 promotes DNA uptake by DCs independently of TLR2, TLR4, RAGE or CD24, suggesting other receptor(s) mediate the process of HMGB1-dependent DNA internalization. Further, DNA-HMGB1 induced lysosomal membrane destabilization which may facilitate DNA release into the cytosol for sensing and trigger activation of the cGAS-STING pathway. Citation Format: Daiana P. Celias, Kay Hänggi, Brian Ruffell. Investigating the mechanisms involved in HMGB1-dependent DNA uptake and STING activation in dendritic cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 678.
Abstract 2758: Expanding PRISM high-throughput screening capabilities to other modalities
Authors: Jacob Smigiel, Jillian Eskra, Antonella Masciotti, Aydin Golabi, Blanche Ip, Melissa Ronan, Matthew G. Rees, Jennifer Roth
Journal: Cancer Research (2023)

Abstract PRISM, a high-throughput drug screening platform, has been used extensively to profile DMSO-soluble compounds, including nearly all commercially available FDA-approved small molecule drugs, in pools of cancer cell lines. In addition to these highly-optimized compounds, novel small-molecule libraries comprising tens of thousands of compounds have been screened through collaborative partnerships. While PRISM has been highly effective in discovering new therapeutic targets and classes via libraries of this nature, we have not yet explored screening aqueous molecules at scale, such as antibodies, antibody-drug conjugates (ADCs), cytokines, and other biologics. There is also untapped potential for the further development of PRISM-based assays utilizing more physiologically relevant assay conditions to gain new insights into cancer biology and highlight more clinically relevant targets. Specifically, the incorporation of environmental cues such as media nutrients, extracellular matrix, and other non-cancer cell types can have a drastic impact on cancer cell state, and thus cancer cell responses to therapy. For example, PRISM pools grown as 3D spheroids, compared to 2D mono-layers, reveal models showing increased sensitivity to mutant KRAS inhibition. Similarly, the addition of immune effector cells, via co-culture methods, to the PRISM platform will enable identification of key mechanisms involved in antibody-dependent cellular cytotoxicity and measurement of immunotherapeutic-relevant phenotypes at an unprecedented scale. This coupling of aqueous screening, with the addition of relevant tumor-microenvironmental cues may help identify therapies more destined to succeed in the clinic, and delineate novel mechanisms involved in cancer cell-state that underpin the many clinical challenges we face in curing cancer. Citation Format: Jacob Smigiel, Jillian Eskra, Antonella Masciotti, Aydin Golabi, Blanche Ip, Melissa Ronan, Matthew G. Rees, Jennifer Roth. Expanding PRISM high-throughput screening capabilities to other modalities [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2758.